Rabbit anti p21 10355 1 ap

Western blotting was performed and analyzed as described
[84 (link)]. Primary antibodies used were mouse anti-MYC (9E-10, Santa Cruz Biotechnology), mouse monoclonal anti-CIP2A (2G10, Novus Biologicals), rabbit anti-E6AP (H-182, Santa Cruz Biotechnology), mouse anti-p53 (DO-1, Santa Cruz Biotechnology), rabbit anti-RB1 (10048-2-Ig, Proteintech), rabbit anti-p21 (10355-1-AP, Proteintech), rabbit anti-14-3-3ζ (C-16, Santa Cruz Biotechnology), and mouse anti-tubulin (Sigma-Aldrich).

HCT-8 cells were lysed in RIPA buffer (25 mM Tris-HCl, pH = 7.4; 150 mM NaCl; 1% NP-40; 0.5% Na-deoxycholate) supplemented with protease inhibitor cocktail. Proteins were separated by 10–12% SDS–PAGE, blotted onto PVDF membrane and incubated with primary antibodies (mouse anti‐GAPDH, 60004-1-Ig, Proteintech, 1:5000; rabbit anti-NOP53, CST, 1:1000; rabbit anti-p53, 10442-1-AP, Proteintech, 1:1000; rabbit anti-p21, 10355-1-AP, Proteintech, 1:1000) overnight at 4 °C, followed by incubation with appropriate secondary antibody (Goat Anti-Rabbit IgG(H + L) HRP, GAR0072, MULTISCIENCES and Goat Anti-Mouse IgG(H + L) HRP, GAM0072, MULTISCIENCES, Hangzhou, China, 1:5000) for 1 h at RT. The protein intensity was analyzed using Image Lab v5.2.1.